Emam Merna H, Mahmoud Mohamed I, El-Guendy Nadia, Loutfy Samah A
Nanotechnology Research Center (NTRC), the British University in Egypt, Suez Desert Road, El-Shorouk City, P.O. Box 43, Cairo, 11837, Egypt.
School of Biotechnology, Badr University in Cairo, Badr City, 11829, Cairo, Egypt.
AMB Express. 2024 Sep 16;14(1):104. doi: 10.1186/s13568-024-01739-8.
Developing a potent antiviral agent to combat Coronavirus Disease-19 (COVID-19) is of critical importance as we may be at risk of the emergence of new virus strains or another pandemic recurrence. The interaction between the SARS-CoV-2 spike protein and Angiotensin Converting Enzyme 2 (ACE2) is the main protein-protein interaction (PPI) implicated in the virus entry into the host cells. Spike-ACE2 PPI represents a major target for drug intervention. We have repurposed a previously described protein-protein interaction detection method to be utilized as a drug screening assay. The assay was standardized using Chitosan nanoparticles (CNPs) as the drug and SARS-CoV-2 spike-ACE2 interaction as the PPI model. The assay was then used to screen four natural bioactive compounds: Curcumin (Cur), Gallic acid (GA), Quercetin (Q), and Silymarin (Sil), and their cytotoxicity was evaluated in vitro. Production of the spike protein and the evaluation of its activity in comparison to a standard commercial protein was part of our work as well. Here we describe a novel simple immunofluorescent screening assay to identify potential SARS-CoV-2 inhibitors that could assess the inhibitory effect of any ligand against any PPI.
开发一种有效的抗病毒药物来对抗冠状病毒病-19（COVID-19）至关重要，因为我们可能面临新病毒株出现或另一场大流行复发的风险。严重急性呼吸综合征冠状病毒2（SARS-CoV-2）刺突蛋白与血管紧张素转换酶2（ACE2）之间的相互作用是病毒进入宿主细胞所涉及的主要蛋白质-蛋白质相互作用（PPI）。刺突-ACE2 PPI是药物干预的主要靶点。我们重新利用了一种先前描述的蛋白质-蛋白质相互作用检测方法，将其用作药物筛选测定法。该测定法以壳聚糖纳米颗粒（CNPs）作为药物，以SARS-CoV-2刺突-ACE2相互作用作为PPI模型进行标准化。然后该测定法用于筛选四种天然生物活性化合物：姜黄素（Cur）、没食子酸（GA）、槲皮素（Q）和水飞蓟素（Sil），并在体外评估了它们的细胞毒性。我们的工作还包括刺突蛋白的生产及其与标准商业蛋白相比的活性评估。在此，我们描述了一种新型的简单免疫荧光筛选测定法，以鉴定潜在的SARS-CoV-2抑制剂，该方法可以评估任何配体对任何PPI的抑制作用。
